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Apoptotic sensitivity in osteoblasts is maturation specific
Author(s) -
Mastracco Dominick,
Saunders Ray,
Scheinfeld Victoria,
Adams Christopher S
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.lb31
Subject(s) - apoptosis , osteoblast , in vitro , chemistry , microbiology and biotechnology , mtt assay , andrology , endocrinology , medicine , biology , biochemistry
As the microenvironment of bone contains significant levels of apoptogens, we hypothesize that maturation of osteoblasts is characterized by changes in sensitivity to those apoptogens. Human Fetal Osteoblast (hFOB) cells cease proliferation and differentiate when cultured above 37°C and maintain a proliferative phenotype at 34°C. Thus, these cells can serve as an in vitro model for osteoblast maturation. hFOBs were cultured at 34°C and 39°C and exposed to elevated levels of the phosphate and calcium ion pair for 24 h. Viable cells were identified by MTT assay and apoptosis confirmed using Mitochondrial ToxGlo and Caspase‐3/7 assays. Results indicated that differentiated osteoblasts had an increased sensitivity to the ion pair. The mitochondrial membrane potential rose initially after apoptogen exposure, followed by a loss of potential consistent with a permeability transition event. The caspases were seen to steadily increase following exposure to the apoptogen. These results indicate that osteoblasts have increased sensitivity towards apoptosis with maturation. This study was supported by a grant from the Center for Chronic Disorders of aging at the Philadelphia College of Osteopathic Medicine.